BBa_K2066003

BBa_K2066003 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K2066003
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Kalen Clifton, Christine Gao, Andrew Halleran, Ethan Jones, Likhitha Kolla, Joseph Maniaci, John Marken, John Mitchell, Callan Monette, Adam Reiss
Date created: 2016-08-24 11:00:00
Date modified: 2016-10-12 12:00:17

Tet Monomer B w/ 8 bp spacer for ICA



Types
DnaRegion

Roles
engineered_region

Composite

Sequences BBa_K2066003_sequence (Version 1)

Description

The part is flanked by UNS 2 and UNS 3 as per the W&M UNS gibson cloning standard. This part is the TetO, 8BP spacer 'B' monomer for use in Iterative Capped Assembly to create repeated TetO Arrays of variable length. The monomer can be PCR amplified using UNS 4 and UNS 5 PCR landing pads. Next, the part should cut with BsmBI restriction enzyme to expose sticky ends and used to extend monomer in ICA. Monomer B (this part) can bind to Monomer A (BBa_K2066002) . This part contains the following: UNS 2 (WM standard gibson/amplification primer site) ??? UNS 4 (used to provide orthogonal amplification of monomers alone) ??? BsmBI site ??? sticky end 2 - Tet Repeat ???8 bp spacer (taken as first 8 bp from UNS X) ??? BsmBI site ??? Sticky end 3 - UNS 5 (orthogonal amplification in conjunction with UNS 4) ??? UNS 3 (WM standard gibson/amplification primer site).

Notes

Design of monomer part was based off of TALEN monomer design in Briggs et al., 2012.

Source

ICA method and sequence design based on Briggs et al., 2012 and its supplement.
Briggs, A. W., Rios, X., Chari, R., Yang, L., Zhang, F., Mali, P., & Church, G. M. (2012). Iterative capped assembly: rapid and scalable synthesis of repeat-module DNA such as TAL effectors from individual monomers. Nucleic acids research, gks624.
The UNS2, UNS3, UNS 4, and UNS 5 sequences are taken from Torella et al. 2013 and are ideal for cloning long sequences of monomers.
Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860.

igem#experience
None
 
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K2066003/1